Apparatus and method for a lysis of a sample, in particular for an automated and/or controlled lysis of a sample

ABSTRACT

The present invention provides an apparatus and a method for a lysis procedure, in particular for an automated and/or controlled lysis procedure of a sample, in particular a biological sample. The apparatus comprises at least one rotation disc ( 31 ), at least one vial holder ( 90 ) which is configured to receive a vial ( 100 ), wherein the vial holder ( 90 ) is arranged on the disc ( 31 ), at least one driving device ( 20 ) which is configured to rotate the disc ( 31 ) and the vial holder ( 90 ), at least one heating device ( 60 ) which is configured to heat the sample to a determined incubation temperature, and—at least one control device ( 70 ) which is configured to control the driving device ( 20 ) and/or the heating device ( 60 ) by means of a timing and/or step control, and/or—at least one transmitting device ( 80 ) for inductive coupling for energy and signal transmission, which is configured to transmit the energy for heating to the heating device ( 60 ), and/or—wherein the driving device ( 20 ) is configured to rotate the disc ( 31 ) in a first direction (A 1 ) and/or with a first speed, and to rotate the vial holder ( 90 ) in a second direction (A 3 ) and/or with a second speed. The apparatus and the method are adapted for an (automated) lysis procedure, wherein the lysis can be carried out in a safe, efficient and effective manner.

TECHNICAL FIELD OF THE INVENTION

The invention relates to an apparatus and a method for a lysis of asample, in particular for an automated and/or controlled lysis of asample, in particular a biological sample.

BACKGROUND OF THE INVENTION

A major task in biochemical and molecular diagnostic laboratories issample homogenization, cell and tissue lysis and the mixing of reagents.The lysis is used for subsequent isolation of biomolecules in the fieldof research and development, chemical analysis and diagnostics.

Today, vortexers and bead mills are routinely used to perform thesetasks. Such systems agitate the fluids and sample material in a vial insuch a way that the vials are accelerated and decelerated in arepetitive pattern in one or more dimensions.

For sample homogenization, beads (glass, ceramic or other materials) areadded. Due to the rapid acceleration and deceleration steps, these beadsgenerate a physical impact force on the sample material and ‘grind’ or‘mill’ it into smallest particles that are then suspended in asurrounding fluid (hence the name ‘bead milling’). On a cellular level,cell walls are cracked under such acceleration, deceleration, impact andshear forces if the bead sizes are selected appropriately. This is forexample necessary for DNA tests where access to the DNA embedded incells is needed. Besides milling with a conventional vortexer, the useof bead mills allows the milling of different materials.

There are also some known chemical agents that improve the cell openingprocess.

The combination of chemical or enzymatic lysis steps with mechanicalmilling steps additionally improves the performance of the lysisprocedure. The use of chemical or enzymatical steps require certainincubation temperatures, as well as thorough mixing of the reagentsduring the incubation steps.

Despite the wide usage, the known vortexing and bead milling devices andprocedures in many cases have low efficiency and therefore need asubstantial amount of processing time.

Additionally, these methods need to be combined with incubation steps atdifferent temperatures to allow efficient lysis of a wide variety ofclinical samples. The current methods require a combination of differentsteps involving the steps of mixing, incubation at differenttemperatures, ideally interrupted by e.g. bead milling. Each of thesteps is performed in different instruments (vortexer, heating block,bead mill . . . ) as a combination of manual steps. That is, thesesequential steps require manual work of well trained operators.

In order to avoid the time-consuming operation due to the severalinstruments needed for a lysis process, efforts have been made to reducethe number of the necessary devices. However, with conventional devicesit is not possible to overcome for example problems caused by the highG-forces which occur due to rotation operations (several steps cannot becarried out or cannot be carried out in an appropriate manner during therotation of the vial, for example appropriate heating and/or determiningthe temperature of the sample or even temperature regulation of theagitated sample).

With other known methods, the bead milling step is performed at lowertemperatures. Several of the bead mills commercially available areequipped with a system which allows the sample to be milled in arefrigerated compartment of the instrument. Other systems allow forsubmersing the grinding beaker or vial into liquid nitrogen before beadmilling, so that the process is carried out at low temperatures.However, such devices do not provide a desired degree of efficiency,especially when treating viscous biological samples.

An analysis of mechanical effects during the bead milling process showsthat the efficiency mainly depends on the strength of the impactcollisions of the beads with the suspended sample material and cells.Such collision forces can be increased by increasing the density of thebeads, the amount of beads and/or increasing the acceleration anddeceleration speed. Today's vortexers and bead mills are already attheir limits with respect to acceleration and deceleration speeds due totheir construction.

A typical known system does not only accelerate and decelerate thesample material and vial but also the much larger masses of the vialholders. Furthermore the movements are most often created by a rotatingmotor shaft driving an eccentric tappet. This eccentric tappet thendrives a plate that is spring suspended for example in the x- and z-axis(that is, right and left movements, up and down movements). The plate iscoupled with a vial holder or the vial is pressed on that platemanually. Due to large unbalanced masses, such movements create hugestress on the whole construction, thereby limiting the way of travel(maximum eccentricity/maximum deflection of the vial holder) and themaximum velocities that can be achieved. A typical way of travel is onlya few millimeters with a maximum of 3000 to 4000 cycles per minute. Toavoid that such systems start moving on the desk surface duringoperation, they are often equipped with a very heavy additional mass anddampers in the bottomplate.

SUMMARY OF THE INVENTION

The object or technical problem of the present invention is to providean apparatus and a method for a lysis, in particular for an automatedand/or controlled lysis of a sample, in particular a biological sample,that overcome the above described drawbacks of the conventional devicesand methods. In particular, it is the object of the present invention toprovide an apparatus and a method for a lysis, in particular for anautomated lysis, wherein the lysis can be carried out in a safe,efficient and effective manner.

The above mentioned problems are solved by an apparatus for a lysis, inparticular for an automated lysis of a sample, in particular abiological sample, and by a method for a lysis, in particular for anautomated lysis of a sample, in particular a biological sample.

The present invention will now be described by defining differentaspects of the invention. Each aspect so defined may be combined withany other aspect or aspects unless clearly indicated to the contrary. Inparticular, any feature indicated as being preferred or advantageous maybe combined with any other feature or features indicated as beingpreferred or advantageous.

In a first aspect, the present invention relates to an apparatus for alysis of a sample, in particular for an automated and/or controlledlysis of a sample, wherein the apparatus comprises:

-   -   at least one rotation disc,    -   at least one vial holder which is configured to receive or for        receiving a vial, wherein the vial holder is arranged on the        disc,    -   at least one driving device which is configured to rotate or for        rotating the disc and the vial holder,    -   at least one heating device which is configured to heat or for        heating the sample to at least one determined incubation        temperature,    -   at least one control device which is configured to control or        for controlling the driving device and/or the heating device by        means of a timing and/or step control.

It is possible to provide a cooling device for cooling the sample to atleast one determined temperature. The heating and cooling process can becarried out stepwise.

The wording “apparatus for a lysis of a sample, in particular for anautomated and/or controlled lysis of a sample” is interchangeable withthe wording “apparatus for lysing of a sample, in particular forautomated and/or controlled lysing of a sample”. This applies to allaspects of the present invention.

That is, the control device comprises a timing and/or step (process orsequence) control device or function. Also a step diagram control wouldbe possible with the appropriate components. The at least one controldevice is configured to control the driving device and the heatingdevice, and/or is in communication with the driving device and theheating device, so that the driving device and the heating deviceoperate in a coordinated manner to control the lysis (procedure).Therefore, the steps are carried out in a determined order by means ofthe timing and/or step control.

An essential point of the invention in accordance with the first aspectis that the automated lysis or lysis procedure (with all the necessarysteps like vortexing (that is, in particular mixing), grinding, milling,heating and keeping the vial holder (and therefore the sample) in aresting phase) can be carried out without the need of a well trainedoperator, since the necessary steps are automatically performed bydetermining and/or considering the time flow and/or the course of actionof the procedure. That is, the control device operates the apparatus ina manner that the driving device and the heating device work in acoordinated manner, wherein the control device “decides”, due to time(timing) and/or step control (also feedback control), when the differentsteps of the process (heating or rotating, keeping the vial holder in aresting phase) have to be carried out. Thus, the control device startsand stops the corresponding step, and controls the subsequent use orapplication of different rotation speeds and/or angular velocity andincubation temperatures. That is, the apparatus operates in an automatedmanner and carries out the whole lysis procedure without unnecessaryinterruptions and/or using different devices for the procedure.Additionally, with the control device, a precise temperature regulationwithin the fast rotating grinding stations (vial holders) is provided(the control device allows for carrying out a feedback control).

A combination of appropriate buffers and enzymes with bead milling (inthe apparatus for automated lysis of a sample) at elevated temperaturesand resembling the incubation temperatures of the respective enzymesallows very efficient lysis of different sample materials with anidentical protocol (that is, different sample material can be treated inthe same manner).

All preferred embodiments of the second, the third and the fourth aspectcan also be preferred embodiments with respect to the first aspect (seethe preferred embodiments with respect to the driving device, theheating device, the control device and the transmitting device).

Preferably, the driving device comprises driving means for driving thedisc, a central pin, to which the disc is pivot-mounted, a centralpinion, fixed on the central pin, at least a first stage gear or gearwheel, driven via the disc by the central pinion, and at least a secondstage gear or gear wheel, driven by the first stage gear. The centralpin and therefore the central pinion are preferably non-rotatingelements and are fixed. The sample holder is associated with the secondstage gear, and the first stage gear and the second stage gear arepivot-mounted on or within the disc. The disc and the gear (at least themoveable parts of the gear) defining a rotating device or at least apart of the rotating device. Also the central pin and the central pinioncan be seen as part of the rotating device, although these components donot rotate.

The gear (that is the first stage gear wheel and the second stage gearwheel) is preferably driven via the disc by the fixed pinion in the disccenter. In this case, the disc is a carrier carrying the gear wheels anddue to driving the disc, the gear wheels are driven correspondingly.

Preferably, the heating device comprises at least one heating sleeve(heating element) which provides the necessary incubationtemperature(s). The heating sleeve preferably surrounds the vial holderand/or the second stage gear (preferably, each of the vial holders issurrounded by a heating sleeve). With the heating device a precisetemperature regulation/controlling within the fast rotating vial holderis possible.

It is important to balance the thermal mass of the heating sleeve in away that it is big enough to allow efficient and homogenous temperaturetransfer to the vial holder (and therefore to the sample over the vial),and small enough to allow efficient cooling down of the sample withinthe vial holder by convective cooling.

Preferably, the heating device comprises or is associated with a leastone temperature sensor. The sensor is associated with the heatingsleeve, the vial holder and/or the sample, which detects the temperatureof the heating sleeve, the vial holder and/or the sample and provides acorresponding signal (temperature detecting sensor).

As already stated above, the control device is configured to control thedriving device and/or the heating device, so that the driving device andthe heating device operate in a coordinated manner to control the lysisprocedure or lysis steps.

The control device preferably comprises a time and/or step controldevice or function, in particular a feedback control device, or isconfigured to carry out a time and/or step control, in particular afeedback control, for controlling the driving device and/or the heatingdevice. That is, the progress or course of action of the single steps ofthe lysis procedure is controlled by means of the time and/or stepcontrol device or function and therefore, by means of the controldevice.

The control device preferably comprises a driving control device orfunction, and/or a timing control device or function, and/or atemperature control device or function for controlling the drivingdevice and/or the heating device. These devices are configured to carryout the time and/or step control (the devices are in communication withthe driving device and the heating device to carry out the time and/orstep control).

Preferably, the control device is configured to control the drivingdevice and/or the heating device at least in response to the signal(s)provided by the temperature sensor(s).

The control device is for examples configured as the time and/or stepcontrol device which control(s) the single steps necessary for carryingout the lysis procedure. The control device (or devices) initiates(starts) and stops the single steps at a desired or determined point oftime (determines and/or adjusts the duration of the single steps),determines and/or adjusts the direction and the speed of the rotation ofthe components (for example of the disc) and determines and/or adjuststhe temperatures which have to be achieved (incubation temperatures).Due to feedback, for example due to a measured temperature and/or due tothe expiry of a required or determined time period, the course of action(the following steps) are carried out in the required manner.

With an ideal protocol (example), the following steps are for examplecarried out:

-   a) 1 minute milling the sample (rotating phase) during heating up    the sample to the first determined incubation temperature, for    example 56° C.;-   b) 5 minutes keeping the sample unmoved (resting phase) at the first    determined incubation temperature;-   c) 0.5 minutes mixing the sample (rotating phase) at the first    determined incubation temperature;-   d) 5 minutes keeping the sample unmoved (resting phase) at the first    determined incubation temperature;-   e) heating up the sample (resting phase) to the second determined    incubation temperature, for example 96° C.;-   f) 5 minutes milling the sample (rotating phase) at the second    determined incubation temperature;-   g) 10 minutes keeping the sample unmoved (resting phase) at the    second determined incubation temperature;-   h) 0.5 minutes cooling down the sample.

A rotating phase is performable or can be carried out for a determinedtime period. A resting phase is a phase in which no rotation is carriedout, that is, an unmoved phase or a phase without any movement orwithout a rotation is provided. Also the resting phase is performable orcan be carried out for a determined time period. The time periods arecontrolled by the control device.

Preferably, the step of rotating comprises rotating the disc in a firstdirection and rotating the vial holder in a second direction, oppositeto the first direction, and while the disc performs one rotation, thevial holder performs one rotation in the opposite direction, thus alwaysor substantially keeping the absolute orientation of the vial holdersubstantially constant while rotating.

The lysis can be carried out for example as follows (as an example forthe steps a) and b)):

When starting the lysis procedure, the driving device starts (therotation of the disc) and for example, the driving control devicecontrols the driving device with respect to the direction of rotation ofthe disc and with respect to the speed, and the timing control devicecontrols the driving device with respect to the duration of the rotatingphase of step a). Additionally, the heating device starts and thetemperature control device controls the heating device, and therefore,the temperature of the sample (by means of the temperature sensor, thatis monitoring the temperature).

The timing control device provides an output signal upon expiry of thetime period of step a) for stopping the driving device (feedback controlby means of monitoring the duration of the time period). The temperaturecontrol device provides an output signal upon receiving a correspondingsignal from the temperature sensor that the determined incubationtemperature has been reached (feedback control by means of the signalsprovided by the temperature sensor).

Then, the next step, step b) can be triggered. The timing control devicethus controls the driving device (resting phase for 5 minutes) andprovides an output signal upon expiry of this time period. The heatingdevice will be controlled to keep the incubation temperature (forexample 56° C.). After expiry of the time period of step b), the nextstep, step c) can be triggered etc. The heating device is configured tocontrol the temperature during a rotating phase and/or during a restingphase.

Preferably, the driving device comprises a planetary gear. That is, theat least first stage gear(s) and the second stage gear(s) are arrangedas part of the planetary gear. The planetary gear allows the drive ofthe disc and the vial holders to be performed in an efficient manner.Additionally, a planetary gear is small in size and thus, the apparatuscan be provided as a compact device.

Preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed and/or (angular) velocity, and    -   to rotate the vial holder in a second direction, and/or with a        second speed and/or (angular) velocity.

Preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed and/or (angular) velocity, and    -   to rotate the vial holder in a second direction, opposite to the        first direction, and/or with a second speed and/or (angular)        velocity.

In principle, it is possible to rotate the disc and the vial holder inthe same direction (that is, the first direction is equal to the seconddirection; it is also possible that the first speed is equal to thesecond speed). However, the best results are achieved when the vialholder rotates relative to the disc.

Preferably, the driving device is configured to rotate the disc and thevial holder such that or in such a manner that the absolute orientationof the vial holder is kept constant while rotating. That is, theabsolute orientation of the vial holder is always kept constant whilerotating.

More preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed and/or (angular) velocity, and    -   to rotate the vial holder in a second direction, opposite to the        first direction, and/or with a second speed and/or (angular)        velocity,        and while the disc performs one rotation, the vial holder        performs one rotation in the opposite direction, thus (always)        keeping the absolute orientation of the vial holder        substantially constant while rotating.

Experimental work showed that further increasing acceleration anddeceleration (heavy deceleration) as well as larger movementssubstantially increase efficiency of the bead milling effect. Theinvention overcomes the major problems as described above by avoidingany unbalanced mass in the system (like in a vortexer), although forexample mixing, milling and grinding steps (that is, also “shaking”steps) are carried out and acceleration is increased. The vial holders,for example two or more, are equally spaced on a circle on the rotatingdisc.

The specific type of movement (for example by means of the planetarygear, which allows for moving the disc and the vial holder in a desireddirection, for example in opposite directions) allows for carrying outall necessary steps for a lysis procedure, that is, vortexing, grinding,and milling the sample. Conventional vortexers merely allow for“shaking”, that is mixing the sample, however no further steps (likeheating) can be carried out. Additionally, vortexers do not run smoothdue to the eccentric arrangement and therefore, there are limitationswith respect to the rotation speed (acceleration). The individualmovement of the disc and the vial holder in accordance with the presentinvention allows for carrying out the necessary steps in a veryefficient manner (smooth running, high rotation speeds and therefore,high efficiency).

Preferably the disc is equipped with at least two vial holders. Thisallows for easily balancing the system and avoiding any mass imbalances.In case of only one vial holder the system can be easily balanced byadding an additional mass with the same weight and distribution as thegear. In a preferred configuration, for example four first stage gearsand four second stage gears are provided on or within the disc androtating around the central pinion.

Preferably, and as already mentioned above (and this is a very effectivemanner), the two stage gears (or for example a two stage gear chain)coupled to the center pinion drive the individual vial holder(s) in sucha way that while the rotation disc performs one rotation, the vialholder(s) perform(s) one rotation in the opposite direction, thus alwaysand/or substantially keeping the absolute orientation of the vialholder(s) substantially constant while rotating (balanced masses, forexample in comparison with a conventional vortexer). That is, thedriving device rotates the disc in a first direction, and the vialholder(s) in a second direction, opposite to the first direction,preferably with the same angular velocity. This is in contrast to aconventional vortexer with the eccentric movements and in contrast to aconventional centrifuge (which does not allow for any “shaking” steps)with fixed coupled vial holders, coupled to the rotating disc. Such asystem without a gear only creates a constant centripetal force pressingthe sample and fluids constantly outwards.

The first stage gear rotate for example clockwise and the second stagegear—and thus also the vial holders—rotate anti-clockwise. As the gearratio overall is preferably 1:1, the vial holders rotate anti-clockwisewith the same angular velocity as the disc rotates clockwise. As aconsequence, the vial holders while moving on a circular track do notmake any rotational movements themselves referred to e.g. the fixedfootplate or bottom side and center pin.

The effective movement is a superimposition of at least two circularmovements. The samples contained in the vials in the vial holderseffectively are moved left/right and for- and backwards. The frequencyis identical to the rotation speed of the disc.

Additionally, due to the possibility to rotate the disc as well as thevial holder or the vial holders (individual rotation of disc and vialholder, realized by the driving device, in particular by means of theplanetary gear or arrangement) in a specific manner, the arrangementruns smoothly and with low vibrations, since the specific movement(rotation) of the components (disc, vial holder) allows for harmonizingmass imbalances, although mixing, milling and grinding steps are carriedout. That is, due to the arrangement of the components of the apparatusand the specific possibility of moving the components (disc, vialholder) by means of the driving device, a system with balanced masses isobtained. In other words, the eccentric motion or movement (necessaryfor example in a vortexer) is converted into rotation, thus, the strongacceleration forces do not longer act on the mechanical components. Theindividual rotation of the disc and the vial holder or vial holdersmeans that the disc and the holders are rotatable in differentdirections (clockwise or counterclockwise) and with different speeds,depending for example on the gear ratio. However, rotation of thecomponents in the same manner (in the same direction and/or with thesame speed) is also possible. Also a rotation of the componentsindependently from one another is possible, with a driving deviceproviding different driving means. However, it is the intention torotate the components in such a manner that a balanced system isprovided.

If e.g. the second stage gear has more teeth or fewer teeth than thecenter fixed pinion, the vial holders in addition perform a rotationalmovement superimposed to the original movement with a rotational speeddetermined by the ratio of the number of teeth of the pinions. That is,if the number of teeth of the central pinion (inner pinion) and thesecond stage gear (outer pinion) in the gear are not equal, the vialholders in addition perform a rotational movement that can help improvemixing of fluids in the vials. These rotations (for example superimposedslow rotation) can be adjusted easily by selecting the appropriate teethratio of the inner and outer pinion.

Due to the balanced masses of such a system, the rotation speed can beeasily increased compared to the current systems. The remainingimbalance of masses can only origin from the sample material and volumesitself. With typical sample volumes of a few milliliters, suchimbalances can only be in the order of a few grams maximum. This can betolerated by the inventive device up to several thousand rpm (rounds perminute).

As the maximum acceleration/deceleration is proportional to the radiusand proportional to the square of the rotational speed of the disc,improvements with respect to the diameter of the disc and the rotationspeed compared to the known vortexing devices in the order of 40 areeasily achievable (e.g. ten times the radius—e.g. 5 mm vs. 5 cm—andtwice the speed—3000 rpm vs. 6000 rpm).

Also more discs can be provided, for example staggered, to increase thenumber of available vial holders for high sample throughputapplications.

The apparatus preferably comprises at least one transmitting device forinductive coupling for energy and/or signal transmission (transmittingdevice, transmitting device for transmitting energy and/or signals),which is configured

-   -   to at least transmit the energy for heating to the heating        device, and/or    -   to at least transmit the signal(s) provided by the temperature        sensor to the control device.

The energy for heating the heating device, that is, the sleeve, ispreferably transmitted by means of the transmitting device (means forinductive coupling for energy and signal transmission, inductivetransducer). The heating device is for example formed as a resistanceheating device. It is also possible to use a Peltier element or similardevices.

Preferably, the signal(s) provided by the temperature sensor are alsotransmitted by the transmitting device (also for example the energy forthe sensor). That is, the values, measured by the sensor are for exampletransmitted to the control device, and thus, the time and/or stepcontrol is carried out accordingly. This allows for an easy way oftransmission, without the need of cable, wires or the like. Theconstruction of a bead milling device containing a heatable vial holderis very complex, due to the high rotation speed of the mill, especiallywithin a planetary bead mill, since there are two rotation axes.However, the use of the inductive transducer allows for carrying out thenecessary lysis steps without the need of using different devices. Thetransmission of energy and signals does not interfere with the rotationof the disc and the vial holders.

Due to the cooperation of the control device, the driving device, theheating device and the sensor(s), an automated lysis procedure can becarried out. The transfer of energy and of measurement signals of atemperature probe only has to take place over one rotation axis insteadof two.

In a second aspect, the present invention relates to an apparatus for alysis of a sample, in particular for an automated and/or controlledlysis of a sample, wherein the apparatus comprises:

-   -   at least one rotation disc,    -   at least one vial holder which is configured to receive or for        receiving a vial, wherein the vial holder is arranged on the        disc,    -   at least one driving device which is configured to rotate or for        rotating the disc and the vial holder,    -   at least one heating device which is configured to heat or for        heating the sample to at least one determined incubation        temperature,        wherein the driving device is configured    -   to rotate the disc in a first direction and/or with a first        speed, and    -   to rotate the vial holder in a second direction and/or with a        second speed.

An essential point of the invention in accordance with the second aspectis in particular to provide a system with balanced masses, as describedwith respect to the first aspect. That is, the apparatus allows formixing the sample (like in a conventional vortexing device) andadditionally, the system runs in a smooth manner (balanced masses, seefirst aspect). The inventive apparatus runs in a smooth manner (like aconventional centrifuge), although steps are carried out which areusually carried out by means of a vortexer (vortexing device).

All preferred embodiments of the first, the third and the fourth aspectcan also be preferred embodiments with respect to the second aspect (seethe preferred embodiments with respect to the driving device, theheating device, the control device and the transmitting device). As tothe further explanations and information with respect to theseembodiments (below), it is referred to the explanations and informationprovided above with respect to the first aspect of the invention.

Preferably, the driving device comprises:

-   -   driving means for driving the disc,    -   a central pin, to which the disc is pivot-mounted,    -   a central pinion, fixed on the central pin,    -   at least a first stage gear, driven via the disc by the central        pinion,    -   at least a second stage gear, driven by the first stage gear,        wherein the vial holder is associated with the second stage        gear.

Preferably, the driving device comprises a planetary gear. That is, thefirst stage gear(s) and the second stage gear(s) are arranged as part ofthe planetary gear.

Preferably, the driving device is configured to rotate the disc and thevial holder such that or in such a manner that the absolute orientationof the vial holder is kept constant while rotating.

Preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed, and    -   to rotate the vial holder in a second direction, opposite to the        first direction, and/or with a second speed,        and while the disc performs one rotation, the vial holder        performs one rotation in the opposite direction, thus keeping        the absolute orientation of the vial holder substantially        constant while rotating. That is, the absolute orientation of        the vial holder is always kept constant while rotating.

Preferably, the heating device comprises at least one heating sleevesurrounding the vial holder and/or the second stage gear.

Preferably, the heating device comprises or is associated with a leastone temperature sensor associated with the heating sleeve, the vialholder and/or the sample, which detects the temperature of the heatingsleeve, the vial holder and/or the sample and provides a correspondingsignal.

Preferably, the apparatus comprises at least one control device which isconfigured to control or for controlling the driving device and/or theheating device by means of a timing and/or step control. That is, thecontrol device comprises a timing and/or step control device orfunction.

The at least one control device is configured to control the drivingdevice and/or the heating device, so that the driving device and theheating device operate in a coordinated manner to control the lysis(procedure). Therefore, the steps are carried out in a determined orderby means of the timing and/or step control.

Preferably, the control device comprises a driving control device orfunction, and/or a timing control device or function, and/or atemperature control device or function for controlling the drivingdevice and/or the heating device.

Preferably, the control device is configured to control the drivingdevice and/or the heating device at least in response to the signal(s)provided by the temperature sensor(s).

Preferably the apparatus comprises at least one transmitting device forinductive coupling for energy and signal transmission, which isconfigured

-   -   to transmit the energy for heating to the heating device, and/or    -   to transmit the signal(s) provided by the temperature sensor to        the control device.

In a third aspect, the present invention relates to an apparatus for alysis of a sample, in particular for an automated and/or controlledlysis of a sample, wherein the apparatus comprises:

-   -   at least one rotation disc,    -   at least one vial holder which is configured to receive a vial,        wherein the vial holder is arranged on the disc,    -   at least one driving device which is configured to rotate the        disc and the vial holder,    -   at least one heating device which is configured to heat the        sample to at least one determined incubation temperature,    -   at least one transmitting device for inductive coupling for        energy and/or signal transmission, which is configured to        transmit the energy for heating to the heating device.

An essential point of the invention in accordance with the third aspectis in particular that at least the energy and/or the signals aretransmitted in an easy manner, wherein no wires or the like are needed.The transmission of energy and signals does not interfere with therotation of the disc and the vial holders.

All preferred embodiments of the first, the second and the fourth aspectcan also be preferred embodiments with respect to the third aspect (seethe preferred embodiments with respect to the driving device, theheating device, the control device and the transmitting device). As tothe further explanations and information with respect to theseembodiments (below), it is referred to the explanations and informationprovided above with respect to the first aspect of the invention.

Preferably, the driving device comprises:

-   -   driving means for driving the disc,    -   a central pin, to which the disc is pivot-mounted,    -   a central pinion, fixed on the central pin,    -   at least a first stage gear, driven via the disc by the central        pinion,    -   at least a second stage gear, driven by the first stage gear,        wherein the vial holder is associated with the second stage        gear.

Preferably, the driving device comprises a planetary gear. That is, thefirst stage gear(s) and the second stage gear(s) are arranged as part ofthe planetary gear.

Preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed, and    -   to rotate the vial holder in a second direction and/or with a        second speed.

Preferably, the driving device is configured to rotate the disc and thevial holder such that or in such a manner that the absolute orientationof the vial holder is kept constant while rotating.

Preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed, and    -   to rotate the vial holder in a second direction, opposite to the        first direction, and/or with a second speed,        and while the disc performs one rotation, the vial holder        performs one rotation in the opposite direction, thus always        keeping the absolute orientation of the vial holder        substantially constant while rotating.

Preferably, the heating device comprises at least one heating sleevesurrounding the vial holder and/or the second stage gear.

Preferably, the heating device comprises or is associated with a leastone temperature sensor associated with the heating sleeve, the vialholder and/or the sample, which detects the temperature of the heatingsleeve, the vial holder and/or the sample and provides a correspondingsignal.

Preferably, the apparatus comprises at least one control device which isconfigured to control or for controlling the driving device and/or theheating device by means of a timing and/or step control. That is, thecontrol device preferably comprises a timing and/or step control deviceor function.

The at least one control device is configured to control the drivingdevice and/or the heating device, so that the driving device and theheating device operate in a coordinated manner to control the lysis(procedure). Therefore, the steps are carried out in a determined orderby means of the timing and/or step control.

Preferably, the control device comprises a driving control device orfunction, and/or a timing control device or function, and/or atemperature control device or function for controlling the drivingdevice and/or the heating device.

Preferably, the control device is configured to control the drivingdevice and/or the heating device at least in response to the signal(s)provided by the temperature sensor(s).

Preferably, the transmitting device for inductive coupling for energyand/or signal transmission is configured to transmit the signal(s)provided by the temperature sensor to the control device.

In a fourth aspect, the present invention relates to an apparatus for alysis of a sample, in particular for an automated and/or controlledlysis procedure of a sample, in particular a biological sample. Theapparatus comprises

-   -   at least one rotation disc,    -   at least one vial holder which is configured to receive a vial,        wherein the vial holder is arranged on the disc,    -   at least one driving device which is configured to rotate the        disc and the vial holder,    -   at least one heating device which is configured to heat the        sample to at least one determined incubation temperature,        and    -   at least one control device which is configured to control the        driving device and/or the heating device by means of a timing        and/or step control, and/or    -   at least one transmitting device for inductive coupling for        energy and/or signal transmission, that is for inductive        coupling of energy and/or signals, which is configured to        transmit the energy for heating to the heating device, and/or    -   wherein the driving device is configured to rotate the disc in a        first direction and/or with a first speed, and to rotate the        vial holder in a second direction and/or with a second speed.

All preferred embodiments of the first, the second and the third aspectcan also be preferred embodiments with respect to the fourth aspect (seethe preferred embodiments with respect to the driving device, theheating device, the control device and the transmitting device). As tothe further explanations and information with respect to theseembodiments (below), it is referred to the explanations and informationprovided above with respect to the first aspect of the invention.

The apparatus contains all essential points or qualities of the aspectsas described above.

Preferably, the driving device comprises:

-   -   driving means for driving the disc,    -   a central pin, to which the disc is pivot-mounted,    -   a central pinion, fixed on the central pin,    -   at least a first stage gear, driven via the disc by the central        pinion,    -   at least a second stage gear, driven by the first stage gear,        wherein the vial holder is associated with the second stage        gear.

Preferably, the driving device comprises a planetary gear. That is, thefirst stage gear(s) and the second stage gear(s) are arranged as part ofthe planetary gear.

Preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed and/or (angular) velocity, and    -   to rotate the vial holder in a second direction, opposite to the        first direction, and/or with a second speed and/or (angular)        velocity.

Preferably, the driving device is configured to rotate the disc and thevial holder such that or in such a manner that the absolute orientationof the vial holder is kept constant while rotating.

Preferably, the driving device is configured

-   -   to rotate the disc in a first direction and/or with a first        speed, and    -   to rotate the vial holder in a second direction, opposite to the        first direction, and/or with a second speed,        and while the disc performs one rotation, the vial holder        performs one rotation in the opposite direction, thus keeping        the absolute orientation of the vial holder constant while        rotating. That is, the absolute orientation of the vial holder        is always kept constant while rotating.

Preferably, the heating device comprises at least one heating sleevesurrounding the vial holder and/or the second stage gear.

Preferably, the heating device comprises or is associated with a leastone temperature sensor associated with the heating sleeve, the vialholder and/or the sample, which detects the temperature of the heatingsleeve, the vial holder and/or the sample and provides a correspondingsignal.

Preferably, the control device comprises a time and/or step controldevice or function for controlling the driving device and/or the heatingdevice.

Preferably, the control device comprises a driving control device orfunction, and/or a timing control device or function, and/or atemperature control device or function for controlling the drivingdevice and/or the heating device.

Preferably, the control device is configured to control the drivingdevice and/or the heating device at least in response to the signalprovided by the temperature sensor.

Preferably, the transmitting device for inductive coupling for energyand/or signal transmission is configured to transmit the signal(s)provided by the temperature sensor to the control device.

The following preferred embodiments are part of all aspects of thepresent invention, in particular of the first, second, third and fourthaspect (and also of the aspects below, the fifth, the sixth, theseventh, the eighth, the ninth, the tenth, the eleventh and the twelfthaspect of the present invention).

The driving means or means for driving the disc preferably comprises amotor and a member which connects the motor with the disc, for example atooth belt. That is, the disc is configured to be driven by means of amotor. Another implementation could be made by building the rotatingdisc in such a way that it can act as the rotor of a motor (adding amagnetic structure). In that case a stator built around the disc willcomplete the motor drive structure. This embodiment has the advantage ofnot needing any belt transmission or any drive gear.

The motor can be part of the apparatus for the automated lysis or can beprovided as an additional component which is arranged outside of theapparatus. Since the apparatus is provided with a gear, one motor issufficient for rotating the disc and the vial holder(s). However, it isalso possible to provide a mechanism with which the disc and the vialholder(s) are driven in a separate manner, for example with separatemotors. In this case, one of the gears (of the first or second stage)can be driven directly by an integrated motor. Also an external motor isusable.

The gears, embedded into the rotating disc, can also be built with moregear stages e.g. with four stages instead of two stages.

Each of the gears is preferably mounted within the disc by means of abearing, preferably by means of a ball bearing, a bush bearing or amagnet bearing. Also the disc is pivot-mounted on the central pin bymeans of a bearing. The bearings reduce the friction, in particularsince high rotation speeds are envisioned.

Further improvements are possible, for example with the use of vialshaving a specific configuration. Improvements in the efficiency of thelysis can be obtained not only when using vials with a circularcross-section, but also when using vials with a non-circularcross-section, in particular with an elliptic cross-section or with anyuneven cross-section. That is, the vial holder should be for exampleformed in a manner that also such vials are receivable. No modificationof the vial holder(s) is necessary, if only the inner shape of a vial isformed in the specific manner as described above.

Vials with a cone-shaped bottom create efficient fluid and beadmovements in an up-down direction.

For use in automated systems that allow loading and unloading of vialsby a handling robot, the driving device, in particular the rotatingdisc, can preferably be equipped with or be associated with at least oneposition sensor allowing to detect exact start/stop positions of thevial holders. The position sensor is configured to detect a startposition and/or a stop position of the disc and/or of the vial holder(s)(start/stop position detecting sensor). It is also possible to transmitthe signals from the at least one position sensor by the means forinductive coupling, that is, the transmitting device.

The position sensors can also be associated with the at least one vialholder. That is, each vial holder can be provided with at least oneposition sensor.

Preferably, the vial holder is arranged substantially perpendicular tothe disc or with an incline with respect to the disc. Therefore, theapparatus preferably comprises a deviation device which allows forarranging the vials inclined with respect to the disc (tangentialdeviation, non-right angle). The deviation device is configured toadjust the position of the vial holder, so that the vial holder isarranged substantially perpendicular to the disc or with an incline withrespect to the disc. Such an arrangement can create a strong up and downmovement of fluids, beads and sample materials during the rotation.

Preferably, the apparatus comprises at least two vial holders which areequally spaced on a circle on the disc. This will allow to easilybalance the system and to avoid any mass imbalances. Providing more thantwo vial holders is also possible.

The incubation temperature is preferably in the range of 50° C. to 100°C., preferably at about 52° C., 53° C., 54° C., 55° C., 56° C., 57° C.,58° C., 59° C. and/or about 93° C., 94° C., 95° C., 96° C., 97° C., 98°C., more preferably at 56° C. and/or 96° C. For example an incubationtemperature of 56° C. is used for the optimization of proteinaseactivity and additionally, an incubation temperature of 96° C. is usedfor the optimization for inactivating of the pathogens (withoutcooking).

The control device is preferably provided with a memory device andpreferably with an input- and display device, so that an operator hasthe possibility to communicate with the apparatus and to input forexample the conditions for the desired time and/or step control. Thecontrol device stores the conditions and runs the apparatus in thedesired manner. The memory device and the input and display device canbe provided as integrated devices (within the control device and/or theapparatus) or as additional devices.

The apparatus preferably comprises a device which allows for anadditional up-down movement of the vial holder.

Due to the absence of strong vibrations, the integration of theindividually rotatable disc and vial holder(s) into automated samplepreparation systems becomes possible. With additional retaining elementson the vial holders it also is possible to mount the device in anyarbitrary position (e.g. vertical).

In a fifth aspect, the present invention relates to a method for/ofperforming a lysis of a sample, in particular an automated and/orcontrolled lysis of a sample, in particular a biological sample, whereinthe method comprises the steps of:

-   -   rotating a disc and a vial holder for milling, grinding and        mixing the sample during a rotating phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase, and    -   controlling the lysis, so that the steps are carried out by        means of a timing and/or step control.

The wording “method for performing a lysis of a sample, in particularfor performing an automated and/or controlled lysis of a sample” isinterchangeable with the wording “method for lysing of a sample, inparticular for automated and/or controlled lysing of a sample”. Thisapplies to all aspects of the present invention.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

In a sixth aspect, the present invention relates to a method forperforming a lysis of a sample, in particular for performing anautomated and/or controlled lysis of a sample, in particular abiological sample, using the apparatus (or with or by means of theapparatus) according to the first aspect of the invention, wherein themethod comprises the steps of:

-   -   rotating the disc and the vial holder for milling, grinding and        mixing the sample during a rotating phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase, and    -   controlling the lysis, so that the steps are carried out by        means of a timing and/or step control.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

In a seventh aspect, the present invention relates to a method forperforming a lysis of a sample, in particular for performing anautomated and/or controlled lysis of a sample, in particular abiological sample, wherein the method comprises the steps of:

-   -   rotating a disc in a first direction and/or with a first speed,        and    -   rotating a vial holder in a second direction and/or with a        second speed,    -   for milling, grinding and mixing the sample during a rotating        phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

In a eighth aspect, the present invention relates to a method forperforming a lysis of a sample, in particular for performing anautomated and/or controlled lysis of a sample, in particular abiological sample, using the apparatus (or with or by means of theapparatus) according to the second aspect of the invention, wherein themethod comprises the steps of:

-   -   rotating the disc in a first direction and/or with a first        speed, and    -   rotating the vial holder in a second direction and/or with a        second speed,    -   for milling, grinding and mixing the sample during a rotating        phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

In a ninth aspect, the present invention relates to a method forperforming a lysis of a sample, in particular for performing anautomated and/or controlled lysis of a sample, in particular abiological sample, wherein the method comprises the steps of:

-   -   rotating a disc and a vial holder for milling, grinding and        mixing the sample during a rotating phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase, and    -   transmitting energy for heating to the heating device.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

In a tenth aspect, the present invention relates to a method forperforming a lysis of a sample, in particular for performing anautomated and/or controlled lysis of a sample, in particular abiological sample, using the apparatus (or with or by means of theapparatus) according to the third aspect of the invention, wherein themethod comprises the steps of:

-   -   rotating the disc and the vial holder for milling, grinding and        mixing the sample during a rotating phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase, and    -   transmitting energy for heating to the heating device.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

In a eleventh aspect, the present invention relates to a method forperforming a lysis of a sample, in particular for performing anautomated and/or controlled lysis of a sample, in particular abiological sample, wherein the method comprises the steps of:

-   -   rotating a disc and a vial holder for milling, grinding and        mixing the sample during a rotating phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase,        and    -   controlling the lysis, so that the steps are carried out by        means of a timing and/or step control, and/or    -   transmitting energy for heating to the heating device, and/or    -   wherein the step of rotating the disc and the vial holder        comprises the steps of:    -   rotating the disc in a first direction and/or with a first        speed, and    -   rotating the vial holder in a second direction and/or with a        second speed,    -   for milling, grinding and mixing the sample during a rotating        phase.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

In a twelfth aspect, the present invention relates to a method forperforming a lysis of a sample, in particular for performing anautomated and/or controlled lysis of a sample, in particular abiological sample, using the apparatus (or with or by means of theapparatus) according to the fourth aspect of the invention, wherein themethod comprises the steps of:

-   -   rotating the disc and the vial holder for milling, grinding and        mixing the sample during a rotating phase,    -   heating the sample to at least one determined incubation        temperature in a rotating phase and/or in a resting phase,    -   maintaining the determined incubation temperature in a resting        phase and/or in a rotating phase,        and    -   controlling the lysis, so that the steps are carried out by        means of a timing and/or step control, and/or    -   transmitting energy for heating to the heating device, and/or    -   wherein the step of rotating the disc and the vial holder        comprises the steps of:    -   rotating the disc in a first direction and/or with a first        speed, and    -   rotating the vial holder in a second direction and/or with a        second speed,    -   for milling, grinding and mixing the sample during a rotating        phase.

In a further step (or in further steps), cooling of the sample to atleast one determined temperature can be carried out.

The wording “method for performing a lysis of a sample, in particularfor performing an automated or controlled lysis of a sample” isinterchangeable with the wording “method for lysing of a sample, inparticular for automated or controlled lysing of a sample”.

All the preferred embodiments of the first aspect of the presentinvention are also part of the other aspects, that is, of the second,the third, the fourth, the fifth, the sixth, the seventh, the eighth,the ninth, the tenth, the eleventh and the twelfth aspect of the presentinvention.

Due to the cooperation of the driving device and the control device, inparticular due to the cooperation of the gear (for example planetarygear) and the time and/or step control, the apparatus is able to performan automated and/or controlled lysis or lysis procedure in an efficient,effective and in a safe manner. Furthermore, due to the determinedsequence control (time and/or step control), and the improved conditions(for example higher acceleration) the high efficiency of the lysis doesnot depend on the composition of the sample. Every kind of sample can beprocessed in an efficient and effective manner.

The driving unit or device is also referred to for example as a discand/or vial holder driving unit or device, the heating device is alsoreferred to for example as a sample heating device, and the control unitis also referred to for example as a driving device and heating devicecontrol unit.

Additional preferred embodiments, advantages and features of the presentinvention are defined in the dependent claims and/or will becomeapparent by reference to the following detailed description and theaccompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a schematic cross-sectional side view of the apparatusaccording to a preferred embodiment of the present invention.

FIG. 2 depicts a schematic cross-sectional view of a disc, a gear and acentral pin (at least a part of a rotating device) of the embodiment ofFIG. 1.

FIG. 3 depicts a perspective schematic view of a part of the rotatingdevice of the embodiment of FIG. 1.

FIG. 4 depicts a further perspective schematic view of a part of therotating device of the embodiment of FIG. 1.

FIG. 5 depicts a perspective schematic sectional view of a part of therotating device, along the line V-V in FIG. 3

FIG. 6 depicts a simplified top view of the driving disc and the gear ofthe embodiment of FIG. 1.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENT

The present invention will now be further described by definingdifferent aspects of the invention generally outlined above in moredetail. Each aspect so defined may be combined with any other aspect oraspects unless clearly indicated to the contrary. In particular, anyfeature indicated as being preferred or advantageous may be combinedwith any other feature or features indicated as being preferred oradvantageous.

The drawings are not necessarily to scale. In certain instances, detailsthat are not necessary for an understanding of the embodiment or thatrender other details difficult to perceive may have been omitted.

The same or equally acting components are provided with the samereference signs.

The term “lysis” describes the disintegration of biological cells toallow access to the cellular constituents.

The term “sample” as used herein includes any reagents, solids, liquids,and/or gases.

The term “cell” includes human, animal and plant cells (includingbacteria and fungi).

FIG. 1 shows schematically and not to scale an apparatus 10 forautomated or controlled lysis of a sample, in particular a biologicalsample. The apparatus 10 allows for sample homogenization, cell andtissue lysis and the mixing of reagents. Therefore, the apparatus 10performs or is configured to carry out steps like vortexing, grindingand milling of the sample within a vial holder and also the step ofheating the sample and keeping the vial holder and therefore the samplein a resting phase.

FIG. 2 depicts a schematic cross-sectional view of a disc 31, a gear 40and a central pin 32 (rotating device 30) of the embodiment of FIG. 1,that is, a part of the device shown in FIG. 1 is depicted in an enlargedview. Vortexing (mixing), grinding and milling steps are carried out bymeans of the rotating device 30.

The lysis apparatus 10 in this preferred embodiment comprises a housing11 including a lid 13 which allows for inserting one or more vials 100(see FIG. 4), containing the sample, into the apparatus 10. The lid isfor example arranged at a top side 12 of the housing 11, or the lid 13forms the top side 12 of the housing 11 (as shown in this embodiment).The lid is configured to be removable, for example by means of a slidingmechanism (thereby using for example a motor). A bottom side 14 isformed as a bottom plate, comprising foot members 17, in order toposition the apparatus 10 on a desired location. The rotation disc 31(see FIG. 3) and four vial holders 90 (see FIG. 4) for receiving vials100 are provided, wherein the vial holders 90 are connected with thedisc 31. The disc 31 and the vial holders 90 are rotatable by means of adriving device 20. Furthermore, a heating device 60 is provided, forheating the samples within the vials 100, received in the vial holder 90(see FIG. 2).

In particular FIG. 2 shows the heating device 60 for heating the samplewithin the vials 100. The heating device 60 comprises a heater 61 whichis configured as a heating sleeve surrounding the vial holder (each ofthe vial holders 90 is surrounded by a heating sleeve). A thermalinsulation member 62 insulates the heater 61 from the environment. Theheating device is associated with at least one sensor 63. The sensor 63is associated with the sleeve 61, the vial holder 90 and/or with thesample, and detects the temperature of the heating sleeve, the vialholder and/or the sample. In this embodiment each vial holder isprovided with one sensor (merely one sensor is shown).

For use in automated systems that allow loading and unloading of vialsby a handling robot, the rotating disc can be equipped with a positionsensor 24 (see FIG. 1) allowing for adjusting, controlling and/ordetecting exact start/stop positions for the vial holders (and/or of thedisc).

The disc 31 is pivot-mounted on the central pin 32, wherein the pin 32as a non-rotating element is fixed with a base member (base plate) 15and a cover member (cover plate) 16, mounted within the housing 11, nearthe bottom side 14 and the top side 12. In this embodiment, the pin 32is configured as an elongated member, arranged perpendicular to thebottom side 14, extending from the bottom side 14 to the top side 12 ofthe apparatus. The disc 31 with the vial holders 90 is provided on afirst end 35 of the central pin or member 32 (in use near the top side12 of the apparatus 10). On a second end 36, opposite the first end 35(in use near the bottom side 14), the pin 32 is connected to the basemember 15.

The driving device 20 comprises driving means 21 for driving the disc31. The driving means 21 include a motor 22—in this embodiment arrangedwithin the apparatus 10—and for example a tooth belt 23 for connectingthe motor 22 with the disc 31.

A sleeve 33, surrounding the pin 32 and pivot-mounted on the pin 32 bymeans of bearings is connected with the disc 31 (supports the disc).Additionally, the sleeve 33 comprises a member 34 for engaging the toothbelt 23, that is, for example a toothed pulley 34. In an alternative, aV-belt and a V-belt pulley can be used. That is, in this embodiment, thebelt 23 does not directly engage the disc 31, but does engage the pulley34 and the sleeve 33 which is connected with the disc 31. The sleeve 33can be built integral with the disc 31 or can be provided as separateelement which is connected to the disc.

As an alternative, it is also possible to arrange the belt for exampledirectly on the disc (the belt engages the disc), for example on thecircumference of the disc.

The driving device 20 furthermore comprises the gear 40, in thisembodiment arranged within the disc 31. The disc 31 therefore defines acarrier plate. The gear 40, for example a planetary gear, comprises (inthis embodiment) a central pinion (central wheel) 41 which is fixed onthe central pin 32 and is therefore a non-rotating element. Furthermore,first stage gears (or gear wheels) 42 and second stage gears (or gearwheels) 43 are provided, wherein the vial holders 90 are associated withor connected with the second stage gears 43. Since this embodimentcomprises four vial holders 90, there are also provide four first stagegears and four second stage gears, which are equally spaced on a circleon the rotating disc 31. The first stage gears 42 are driven via thedisc 31 by the central pinion 41, and the second stage gears 43 aredriven by the first stage gears 42.

The disc 31, the central pin 32 with the sleeve 33, and the gear 40defining a rotating device 30 which is driven by means of the drivingmeans 21. The rotatable components are pivot-mounted by means of one ormore bearings 50 (the Figs. show roller bearings) to reduce the frictionbetween the components.

A control device 70 (see FIG. 1) controls the operation of the drivingdevice 20 and/or the heating device 60 in such a manner that the drivingdevice 20 and the heating device 60 work or operate in a coordinatedmanner, preferably by means of a timing and/or step control. Thus, thelysis procedure runs in an automated manner, since the single steps ofthe lysis (for example vortexing, grinding and/or milling of the samplewithin the vial holder and also the steps of heating the sample(s) andkeeping the vial holder and therefore the sample in a resting phase),are automatically controlled. The control device initiates (starts) andstops the single steps at a desired or determined point of time(determines and/or adjusts the duration of the single steps), determinesand/or adjusts the direction and the speed of the rotation of thecomponents (for example of the disc) and determines and/or adjusts thetemperatures which have to be achieved (incubation temperatures).

The control device 70 is part of the apparatus or can be provided as asingle device being associated with the apparatus and the correspondingcomponents, respectively. In FIG. 1, the control device 70 is indicatedas a black box. The control device 70 comprises for example a drivingcontrol device or function 71, and/or a timing control device orfunction 72, and/or a temperature control device 73 or function forcontrolling the driving device 20 and the heating device 60.

The driving control device controls for example the direction ofrotation of the disc and the speed during the rotation, the timingcontrol device controls for example the duration of the rotating phaseand of the resting phase and the temperature or heating control devicecontrols for example the temperature of the sample, the vial holderand/or the heating element (for example the heating sleeve).

Therefore, the control device can be provided as a timing and/or stepcontrol device or can be provided with this function for controlling thedriving device 20 and the heating device 60.

The control device is advantageously provided with a memory device 74and with an input and/or display device 75 to allow for communicationbetween an operator/user and the apparatus. The control device storesinput data provided by the operator via the input device within thememory device and runs the apparatus in the desired manner. Alsopredetermined control processes or courses can be stored in the memorydevice and can be used during operation. The memory device and the inputand display device can be provided as integrated devices (within thecontrol device and/or the apparatus) or as additional devices.

The energy and/or measurement signal transmission is provided by atransmitting device 80 for inductive coupling for energy and/or signaltransmission, that is, by a transmitting device 80 for transmittingenergy and/or signals, in cooperation with the control device 70. Theenergy for heating the heating sleeve(s) is provided by means of thetransmitting device 80 (inductive coupling), and the energy is forexample used to heat at least one wire associated with each of thesleeves (for example in case of resistance heating). Also the signal ofthe temperature sensor (that is, the measured temperature, measured bythe temperature sensor) is transmitted to the control device 70 by meansof the transmitting device 80 (due to inductive coupling) for thefurther control procedure, that is, for example feedback control.

FIG. 3 depicts a perspective schematic view of at least a part of therotating device of the embodiment of FIG. 1. The Fig. shows the disc 31with the transmitting device 80 (means for inductive coupling for energyand signal transmission, inductive transducer) which comprises arotating part 81, connected with the disc 31 and a fixed part 82.

The apparatus furthermore comprises for example a deviation device 78(see FIG. 2) which allows for arranging the vials inclined with respectto the disc (tangential deviation, non-right angle, tilting the axis ofthe vial holder relative to the axis of the disc 31). The device ispreferably integrated within the apparatus, that is, part of theapparatus.

FIG. 4 depicts a further perspective schematic view of at least a partof the rotating device 30 of the embodiment of FIG. 1, without the disc31. Merely the transmitting device 80 is shown. Therefore, the gear 40can be seen. The four vial holders 90 are arranged above the four secondstage gear wheels 43. One of the vial holders 90 contains a vial 100.

FIG. 5 depicts a perspective schematic sectional view of a part of therotating device 30, along the line V-V in FIG. 3.

Having described the main structural features of the apparatus 10 forautomated lysis of a sample, in particular a biological sample accordingto the present invention, the below will describe under furtherreference to FIG. 6 (simplified top view of the rotating disc 31 and thegear 40 of the embodiment of FIG. 1) the function of the apparatus 10during a lysis procedure. The apparatus 10 allows for samplehomogenization, cell and tissue lysis and the mixing of reagents.

In order to perform the lysis of a sample, the sample has to be filledinto a vial, which is configured to be received in the vial holder 90.In the embodiment described with the FIGS. 1 to 5, four vials can beinserted into the four vial holders 90.

The motor 22 drives the disc 31 via the tooth belt 23 and the sleeve 33with the pulley 34 pivot mounted on the central pin 32 for example in aclockwise direction A1 (first direction). The rotation of the disc 31now drives the gear 40 via the central non-rotating pinion, in such amanner that the first stage gears 42 also rotate in the clockwisedirection A2 (first direction). Thus, the second stage gears 43 rotatein a counterclockwise direction A3 (second direction). Rotating the gearwheels in an opposed direction allows for a very efficient and effectivelysis procedure, in particular with the heating of the sample.

The control device 70 controls the driving device 20 for example suchthat while the rotation disc 31 performs one rotation, the vialholder(s) 90 perform(s) one rotation in the opposite direction, thusalways keeping the absolute orientation of the vial holder(s) 90substantially constant while rotating (balanced masses in comparisonwith a conventional vortexer or vortexing device). Due to thesuperimposed circular movements of the disc 31 and the vial holders 90in that specific manner, the system is balanced and runs in a verysmooth manner.

If e.g. the second stage gear has more teeth or fewer teeth than thecenter fixed pinion, the vial holders in addition perform a rotationalmovement superimposed to the original movement with a rotational speeddetermined by the ratio of the number of teeth of the pinions. Such asuperimposed (for example slow) rotation may help mixing the sample andfluids in the vials even better.

The gear ratio and therefore the speed of rotation and the direction ofrotation (of the disc and the vial holder(s)) is definable in a desiredmanner, for example by means of the control device 70. Also other wheelscan be used with another teeth ratio, or the gear can be provided withmore stages. Other kinds of gears are also applicable.

Energy for heating the sleeve(s) 61 is transmitted by the transmittingdevice 80, that is, by means of inductive coupling (energy and signaltransmission). The sleeve is for example configured as resistanceheating, and the required energy is transmitted by the transmittingdevice 80. The transmitting device 80 for inductive coupling is alsoconfigured to allow for transmission of the signals, in particular fortransmission of the temperature value determined by the temperaturesensor(s). Thus, the control device, receiving the measured values fromthe sensor(s), controls the lysis procedure at least by means of themeasured temperature value(s), see also the above provided example of aprotocol with the steps a) to h). Therefore, due to the feedback a timeand/or step control is carried out.

The controllable driving device 20 and the controllable heating device60 allow for an automated lysis procedure by means of the apparatus 10described above. Since the control device 70 operates the apparatus 10by means of a time and/or step control, the single steps of vortexingand mixing, respectively, grinding, milling, heating and keeping thevial holder (and thus, the sample) in a resting phase, the adjustment ofrotation speed, rotation direction and temperature and the adjustment ofthe duration of each step are carried out by means of a defined and/ordetermined (or predetermined) schedule or workflow. Therefore, the lysisis executed automatically, without the need of interruptions and withoutthe need of well trained users controlling the several steps.

Since the apparatus is configured in the above described manner(balanced masses) higher velocities and an increased acceleration arepossible (in comparison with conventional devices). Due to thetransmitting device, energy and/or signals can be transmitted in adesired manner without the need of wires etc. Therefore, the highrotation speeds can be carried out.

The invention can combine the following aspects:

-   -   providing a control device which is configured to control the        steps of the lysis by means of a timing and/or step control;    -   providing a specific arrangement of the apparatus, which allows        for a specific movement of the vial holder(s) and thus,        providing a system with balanced masses;    -   providing a transmitting device (means for inductive coupling)        for energy and/or signal transmission.

The above described invention can be used for any application wherebiological or other material samples (chemistry, food processing, . . .) need to undergo a bead milling procedure to homogenize the materialand/or to effectively mix fluids or suspensions of highly differentviscosity. In particular, the invention is used to build more effectiveand more robust bead mills and vortexers.

LIST OF REFERENCE SIGNS

-   10 apparatus for (automated and/or controlled) lysis-   11 housing-   12 top side-   13 lid-   14 bottom side-   15 base member, base plate-   16 cover member, cover plate-   17 foot member-   20 driving device-   21 driving means-   22 motor-   23 tooth belt-   24 position sensor-   30 rotating device-   31 disc-   32 central pin-   33 sleeve-   34 pulley-   35 first end of the pin-   36 second end of the pin-   40 gear-   41 central pinion-   42 first stage gear-   43 second stage gear-   50 bearing, roller bearing-   60 heating device-   61 heater, heating sleeve-   62 insulation member-   63 temperature sensor-   70 control device-   71 driving control device-   72 timing control device-   73 heating or temperature control device-   74 memory device-   75 input/display device-   78 deviation device-   80 transmitting device-   81 rotating part-   82 fixed part-   90 vial holder-   100 vial-   A1 first direction (disc)-   A2 first direction (first stage gear)-   A3 second direction (second stage gear)

The invention claimed is:
 1. An apparatus for a lysis of a sample, theapparatus comprising: at least one rotation disc, at least one vialholder which is configured to receive a vial, wherein the vial holder isarranged on the disc, at least one driving device which is configured torotate the disc in a first direction and to rotate the vial holder in asecond opposite direction; wherein the driving device comprises a motorfor rotating the disc in the first direction, at least a first stagegear that is rotated by the disc in the first direction, and at least asecond stage gear that is rotated by the first stage gear in the secondopposite direction such that the vial holder that is associated with thesecond stage gear rotates in the second opposite direction; and whereinan absolute orientation of the vial holder is substantially keptconstant while rotating; at least one heater which is configured to heatthe sample to a determined incubation temperature; and at least onecontrol device in communication with and configured to control thedriving device, or the heater, or both the driving device and the heateraccording to input data and a protocol to be carried out automaticallyaccording to at least one of time control, step control, or feedbackcontrol, wherein feedback control is based on a signal received from atleast one of the driving device or the heater, the control devicecomprising: a memory device to store the input data and the protocol;and an input device in communication with the memory device to input theinput data into the memory device.
 2. The apparatus of claim 1, whereinthe driving device further comprises: a central pin, to which the discis pivot-mounted; and a central pinion, fixed on the central pin,wherein at least the first stage gear is driven via the disc by oraround the central pinion.
 3. The apparatus of claim 2, wherein each ofthe gears is mounted within the disc via a bearing.
 4. The apparatus ofclaim 3, wherein the bearing is a ball bearing, a bush bearing, or amagnet bearing.
 5. The apparatus of claim 1, wherein the heatercomprises at least one heating sleeve surrounding the vial holder, orthe second stage gear, or both the vial holder and the second stagegear.
 6. The apparatus of claim 1, wherein: the heater comprises or isassociated with at least one temperature sensor associated with theheating sleeve, the vial holder, or the sample, or a combinationthereof; wherein the temperature sensor detects a temperature of theheating sleeve, the vial holder, or the sample, or a combinationthereof; and wherein the temperature sensor provides a signalcorresponding to the temperature.
 7. The apparatus of claim 6, whereinthe control device controls the heater based on the temperature of theheating sleeve, the vial holder, or the sample, or a combinationthereof.
 8. The apparatus of claim 1, wherein the control device isconfigured to control the driving device or the heater, or both thedriving device and the heater at least in response to the signalprovided by a temperature sensor.
 9. The apparatus of claim 1, whereinthe driving device comprises a planetary gear.
 10. The apparatus ofclaim 1, wherein the driving device is configured to rotate the disc ata first speed and to rotate the vial holder at a second speed.
 11. Theapparatus of claim 1, wherein the apparatus comprises at least oneinductive transducer for inductive coupling for energy, or signaltransmission, or both inductive coupling for energy and signaltransmission, the inductive transducer being configured to transmit atleast one of: energy for heating to the heater or a signal provided by atemperature sensor to the control device.
 12. The apparatus of claim 1,wherein the vial holder is configured to receive a sample vial with acircular cross-section or a sample vial with a non-circularcross-section.
 13. The apparatus of claim 12, wherein the non-circularcross-section is an elliptic cross-section or an uneven cross-section.14. The apparatus of claim 1, wherein the driving device comprises or isassociated with at least one position sensor which is configured todetect a start position, or a stop position, or both the start positionand the stop position of the at least one vial holder, or the disc, orboth the at least one vial holder and the disc.
 15. The apparatus ofclaim 1, wherein the vial holder is substantially perpendicular to thedisc or is inclined with respect to the disc.
 16. The apparatus of claim1, wherein at least two vial holders are equally spaced on a circle onthe disc.
 17. The apparatus of claim 1, wherein the incubationtemperature is in the range of 50° C. to 100° C.
 18. The apparatus ofclaim 17, wherein the incubation temperature is 52° C., 53° C., 54° C.,55° C., 56° C., 57° C., 58° C., 59° C., 93° C., 94° C., 95° C., 96° C.,97° C., or 98° C.
 19. The apparatus of claim 17, wherein the incubationtemperature is 56° C. or 96° C.
 20. The apparatus of claim 1, whereinthe heater comprises at least one temperature sensor and feedbackcontrol is based on a signal received from the temperature sensor. 21.The apparatus of claim 1, wherein the protocol comprises a rotatingphase and a resting phase.
 22. The apparatus of claim 21, wherein therotating phase comprises rotating the rotation disc and the vial holderfor milling, grinding, and mixing the sample.
 23. The apparatus of claim21, wherein the protocol comprises heating the sample to the determinedincubation temperature in the rotating phase.
 24. The apparatus of claim23, wherein the protocol comprises maintaining the determined incubationtemperature in the rotating phase.
 25. The apparatus of claim 21,wherein the protocol comprises heating the sample to the determinedincubation temperature in the resting phase.
 26. The apparatus of claim25, wherein the protocol comprises maintaining the determined incubationtemperature in the resting phase.